Fitoterapia 71 Ž2000.

183]186

Short report

Cytotoxicity and antileishmanial activity of

Annona muricata pericarp
M.C. Jaramillo a , G.J. Arango a,U , M.C. Gonzalez
´ b, S.M.
Robledo c , I.D. Velez c
a
´
Facultad de Quımica ´
Farmaceutica, Uni¨ ersidad de Antioquia, A. A. 1226 Medellın,
´ Colombia
b
Department of Farmacology, Uni¨ ersidad de Valencia, 46100 Burjasot, Spain
c
Facultad de Medicina (PECET), Uni¨ ersidad de Antioquia, A. A. 1226 Medellın,
´ Colombia

Received 16 March 1999; accepted in revised form 18 September 1999

Abstract

Hexane, ethyl acetate and methanol extracts of Annona muricata pericarp were tested in
vitro against Leishmania braziliensis and L. panamensis promastigotes, and against cell line
U 937. The ethyl acetate extract was more active than the other extracts and even of
Glucantime W used as reference substance. Its fractionation led to the isolation of three
acetogenins } annonacin, annonacin A and annomuricin A. Q 2000 Elsevier Science B.V.
All rights reserved.

Keywords: Annona muricata; Acetogenins; Antileishmanial activity; Cytoxicity

´
Plant. Annona muricata L. ŽAnnonaceae., dried pericarp, collected in Medellın,
Colombia, in August 1996 and identified by A. Cogollo ŽHUA.. A voucher
specimen ŽNo. AC 013259. is kept at the Herbarium of University of Antioquia.

Uses in traditional medicine. Rural inhabitants use the pericarp to treat chronic
ulcers w1]4x.

U
Corresponding author.
E-mail address: gjarango@quimbaya.udea.edu.co ŽG.J. Arango A..

0367-326Xr00r$ - see front matter Q 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 3 6 7 - 3 2 6 X Ž 9 9 . 0 0 1 3 8 - 0
184 M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186

Previously isolated classes of constituents. No report.

New-isolated constituents. Annonacin Ž0.0032%., annonacin A Ž0.0021. and anno-

muricin A Ž0.0021. w5]10x from the EtOAc extract.

Annonacin, annonacin A RsH

Annomuricin A R s OH
Annomuricin A. IR bands ŽCHCl 3 .: 1750 cmy1 ŽC s O. and 3400 ŽOH stretch .;
1
H-NMR Ž250 MHz, CDCl 3 .: d 3.8 Ž1H, m, H-4., 3.4 Ž1H, m, H-10., 3.4 Ž1H, m,
H-11., 3.35 Ž1H, m, H-15., 3.8 Ž1H, m, H-16., 1.58 Ž2H, m, H-17., 1.97 Ž2H, m,
H-18., 3.8 Ž1H, m, H-19., 3.89 Ž1H, m, H-20., 7.19 Ž1H, d, H-33., 5.0 Ž1H, m, H-34.,
1.58 Ž3H, d, H-35.; 13 C-NMR: 174.8 ŽC-1., 69.65 ŽC-4., 74.30 ŽC-10., 74.3 ŽC-11.,
74.39 ŽC-15., 82.35 ŽC-16., 28.75 ŽC-17., 28.75 ŽC-18., 82.28 ŽC-19., 71.15 ŽC-20.,
152.15 ŽC-33., 78.12 ŽC-34., 19.0 ŽC-35.; FABMS MHq 613; loss of H 2 O at m r z
595, 577, 559, 541, and 523 indicates the existence of five OH groups. The positions
of the OH groups are suggested by the EIMS ions at m r z 199, 269, 271, 241 and
141. The stereochemistry of the mono THF ring is threo-trans-erythro.

Annonacin. 1 H-NMR: d 3.5 Ž1H, m, H-10., 1.5 Ž2H, m, H-11., 3.4 Ž1H, m, H-15.,
1.61 Ž2H, m, H-17., 3.41 Ž1H, m, H-19., 3.85 Ž1H, m, H-20., 1.42 Ž3H, d, H-35.;
13
C-NMR: 174.0 ŽC-1., 131.0 ŽC-2., 69.83 ŽC-4., 71.51 ŽC-10., 38.0 ŽC-11., 74.02
ŽC-15., 82.48 ŽC-16., 28.37 ŽC-17., 28. 37 ŽC-18., 82.48 ŽC-19., 74.02 ŽC-20., 151.81
ŽC-33., 77.94 ŽC-34.. MHq FABMS 597; loss of H 2 O at m r z 309, 291, 379, 361.
The stereochemistry is threo-trans-threo.

Annonacin A. 1 H-NMR: d 1.2 Ž2H, m, H-11., 3.41 Ž1H, m, H-15., 1.63 Ž2H, m,
H-17., 3.82 Ž1H, m, H-20., 7.17 Ž1H, d, H-33., 1.4 Ž3H, d, H-35.; 13 C-NMR: 69.85
ŽC-4., 71.69 ŽC-10., 74.49 ŽC-15., 82.16 ŽC-16., 83.19 ŽC-19., 74.26 ŽC-20., 151.89
ŽC-33., 79.29 ŽC-34.. The stereochemistry is threo-trans-erythro.

Tested material. Hexane Ž0.89%., EtOAc Ž0.58. and MeOH Ž12.2. extracts.

Used microorganisms. Cell line U-937 was cultured in RPMI 1640 medium supple-
mented with 10% bovine foetal serum and incubated at 378C and 5% CO 2
atmosphere w11x. Leishmania (¨ iannia) braziliensis ŽMHOMrCOr88rUA-301. and
 M.C. Jaramillo F. et al. r Fitoterapia 71 (2000) 183]186 185

Table 1
Antileishmanial activity and cytotoxicity of the extracts from Annona muricata pericarp a

Tested material MEC Žmgrml.

L. (¨ ) braziliensis L. (¨ ) panamensis Cell line U-937

MeOH extract ) 1.0 1.0 ) 1.0

Hexane extract ) 1.0 1.0 1.0
EtOAc extract 0.1 1.0 0.1
Glucantime W 4.1 3.7 0.46
a
MEC, minimum effective concentration.

L. (¨ ) panamensis ŽMHOMrCOr87rUA-140. were isolated from patients from the

north-west of Colombia with cutaneous injuries, conserved in liquid nitrogen, and
cultured in liquid medium ŽSchneider’s insect medium, supplemented with 10%
bovine foetal serum. at 268C.

Studied activity. Antileishmanial activity and cytotoxicity w11x. Tubes were seeded
with either liquid medium with 10 6 promastigotesrml or medium containing 10 6
cell U-937rml, respectively. The minimum effective concentration ŽMEC. was
determined after 48 h of incubation at 378C, using Glucantime W Ž N-methylgluca-
mine antimonate. as a reference substance w3,4x.

Results. Reported in Table 1.

Conclusions. The crude ethyl acetate extract of Annona muricata pericarp was
more active than Glucantime W against both tested strains of Leishmania pro-
mastigotes and cell line U-937. These results, showing in particular an interesting
antileishmanial activity, confirm the importance of the investigation on the thera-
peutic potential of plants used by rural communities.

Acknowledgements

Colciencias, an organization whose objective is to promote scientific and techno-

logical development Colombia, financially supported this work. We wish to thank
Dr Diego Cortes of the Faculty of Pharmacy, Universidad de Valencia ŽSpain..

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